摘要
通过检测椿皮提取物对宫颈癌Hela细胞活力的影响,观察药物对细胞凋亡、细胞周期、细胞自噬及相关因子的调控作用,对引起细胞凋亡的分子机制进行初步研究。体外培养Hela细胞,以不同质量浓度的椿皮提取物处理宫颈癌Hela细胞后,MTT法检测药物对Hela细胞增殖的影响;Annexin V-FITC/PI双染流式细胞术检测椿皮提取物对Hela细胞凋亡的影响;PI单染流式细胞术检测椿皮提取物对Hela细胞周期的影响;qRT-PCR检测椿皮提取物对Bcl-2/Bax mRNA水平表达的影响;Western-blot法测定Hela细胞凋亡因子Bcl-2、Bax及自噬相关因子蛋白的表达。MTT结果显示,椿皮提取物能抑制宫颈癌Hela细胞增殖,呈剂量依赖性(P<0.05);质量浓度为5,10,20μg/mL椿皮提取物处理Hela细胞48 h后,Hela细胞增殖抑制率及凋亡率明显上升,并且高于对照组(P<0.05);PI单染流式细胞术结果发现,椿皮提取物诱导阻滞Hela细胞分裂G2/M期(P<0.05);qRT-PCR结果显示,Bax mRNA表达量升高,Bcl-2 mRNA表达量降低,呈剂量依赖性(P<0.05);Western-blot结果显示,Bax蛋白表达量升高,Bcl-2蛋白表达量降低,LC3BⅡ/Ⅰ值升高,呈剂量依赖性(P<0.05)。椿皮提取物能明显抑制宫颈癌Hela细胞的增殖,通过调控Bax、Bcl-2凋亡基因表达,阻滞细胞G2/M期,促进细胞发生凋亡,通过调控LC3B基因表达,促进宫颈癌细胞发生自噬。
By detecting the effects of Ailanthus altissim bark extract on the viability of cervical cancer Hela cells,and observing the regulatory effects of drugs on apoptosis,cell cycle,autophagy and related factors,we preliminarily studied the molecular mechanism of cell apoptosis.Hela cells were cultured in vitro,and cervical cancer Hela cells were treated with different mass concentrations of Ailanthus altissima bark extract.MTT method was used to detect the effect of drugs on the proliferation of Hela cells;Annexin V-FITC/PI double staining flow cytometry was used to detect the effect of Ailanthus altissima bark extract on Hela cell apoptosis;PI single staining flow cytometry was used to detect the effect of Ailanthus altissima bark extract on the cell cycle of Hela;qRT-PCR was used to detect the effect of Ailanthus altissima bark extract on the expression of Bcl-2/Bax mRNA;Western-blot method was used to determine the expression of Hela cell apoptosis factors Bcl-2,Bax and autophagy-related factor proteins.MTT results showed that Ailanthus altissima bark extract could inhibit the proliferation of cervical cancer Hela cells in a dose-dependent manner(P<0.05);After treating Hela cells with the mass concentration of 5,10,and 20μg/mL Ailanthus altissima bark extract for 48 hours,the proliferation inhibition rate and apoptosis rate of Hela cells increased significantly,and were higher than those of the control group(P<0.05);The results of PI single staining flow cytometry showed that Ailanthus altissima bark extract induced block Hela cell division in G2/M phase(P<0.05);The results of qRT-PCR showed that Bax mRNA expression increased,and Bcl-2 mRNA expression decreased in a dose-dependent manner(P<0.05);Western-blot results showed that Bax protein expression increased,Bcl-2 protein expression decreased,and LC3 BⅡ/Ⅰvalue increased in a dose-dependent manner(P<0.05).Ailanthus altissima bark extract can significantly inhibit the proliferation of cervical cancer Hela cells,by regulating the expression of Bax and Bcl-2 apoptotic genes,blocking the G2/M phase of cells,promoting cell apoptosis,and promoting autophagy in cervical cancer cells by regulating the expression of LC3 B gene.
作者
陈艳艳
于波
王思明
张婉迎
李修刚
李银清
CHEN Yanyan;YU Bo;WANG Siming;ZHANG Wanying;LI Xiugang;LI Yinqing(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China;Jilin Province Zhengkai Pharmaceutical Chain Co.,Ltd.,Changchun 130117,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2021年第6期673-678,共6页
Journal of Jilin Agricultural University
基金
吉林省科技发展计划项目(20190101010JH)。
关键词
椿皮提取物
宫颈癌
HELA细胞
细胞凋亡
Ailanthus altissima bark extract
cervical cancer
Hela cell
cell apoptosis
